SI2726883T1 - A cell mediated immune response assay with enhanced sensitivity - Google Patents

A cell mediated immune response assay with enhanced sensitivity Download PDF

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SI2726883T1
SI2726883T1 SI201231303T SI201231303T SI2726883T1 SI 2726883 T1 SI2726883 T1 SI 2726883T1 SI 201231303 T SI201231303 T SI 201231303T SI 201231303 T SI201231303 T SI 201231303T SI 2726883 T1 SI2726883 T1 SI 2726883T1
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Jeff Boyle
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Cellestis Limited
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5047—Cells of the immune system
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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    • Y02A90/10—Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

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Abstract

This disclosure relates generally to the field of immunological-based diagnostic assays including an assay to measure cell-mediated immunoresponsiveness. The present disclosure teaches diagnosis of a subject's exposure to an antigen based on cell-mediated immunoresponsiveness with enhanced sensitivity. The assay contemplated herein is capable of integration into standard pathology architecture to provide a diagnostic reporting system and to facilitate point of care clinical management.

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Claims (13)

Test celično posredovanega imunskega odziva s povečano občutljivostjo Patentni zahtevkiCell-mediated immune response test with increased sensitivity Patent claims 1. Postopek za merjenje aktivnosti celično posredovanega imunskega odziva v subjektu, pri čemer navedeni postopek obsega so-inkubacijo limfocitov iz subjekta s kombinacijo dveh nizov peptidov, pri čemer prvi niz obsega peptide z dolžino od 7 do 14 aminokislinskih ostankov, ki jih prepoznavajo CD8+ limfociti, in drugi niz, obsega peptide z od 16 aminokislinskih ostankov ali več, ki jih prepoznavajo CD4+ limfociti, pri čemer ti peptidi obsegajo ves ali del proteinskega antigena, in merjenje prisotnosti ali zvišanja nivoja imunske efektorske molekule iz imunskih celic, pri čemer je prisotnost ali nivo imunske efektorske molekule indikativen za nivo celično posredovane odzivnosti subjekta.A method for measuring the cell-mediated immune response activity in a subject, said method comprising co-incubating lymphocytes from a subject by a combination of two strands of peptides, the first strand comprising peptides having a length of from 7 to 14 amino acid residues recognized by CD8 + lymphocytes , and the second strand comprises peptides of 16 amino acid residues or more that are recognized by CD4 + lymphocytes, these peptides comprising all or part of a protein antigen, and measuring the presence or elevation of the immune effector molecule level from immune cells, the presence of or the level of the immune effector molecule indicative of the level of cell mediated response of the subject. 2. Postopek po zahtevku 1, kjer je subjekt človek.The method of claim 1, wherein the subject is a human. 3. Postopek po zahtevku 1, kjer so limfociti iz subjekta vsebovani v krvnem vzorcu in je vzorec nerazredčena polna kri, ali kjer je vzorec polna kri, ki obsega od 10 vol. % do 100 vol. % vzorca, ki naj se testira, zlasti kjer polna kri obsega od 50 vol. % do 100 vol. % vzorca, ki naj se testira, še zlasti, kjer polna kri obsega od 80 vol. % do 100 vol. % vzorca, ki naj se testira.The method of claim 1, wherein the lymphocytes from the subject are contained in the blood sample and the sample is undiluted full of blood, or where the sample is full of blood, comprising 10 vol. % to 100 vol. % of the sample to be tested, in particular where the full blood comprises 50 vol. % to 100 vol. % of the sample to be tested, especially where the full blood comprises 80 vol. % to 100 vol. % of the sample to be tested. 4. Postopek po zahtevku 3, kjer je polna kri zbrana v epruveti, ki obsega heparin.The method of claim 3, wherein the full blood is collected in a tube comprising heparin. 5. Postopek po zahtevku 1, kjer je imunska efektorska molekula citokin, zlasti, kjer je citokin IFN-γ.The method of claim 1, wherein the immune effector molecule is cytokine, in particular, wherein the cytokine is IFN-γ. 6. Postopek po zahtevku 1, kjer se imunski efektorji detektirajo s protitelesi, specifičnimi za le-te, zlasti, kjer se imunski efektorji detektirajo z uporabo ELISA, še zlasti, kjer se imunski efektorji detektirajo z uporabo ELISpot.The method of claim 1, wherein the immune effectors are detected by antibodies specific to these, in particular, where the immune effectors are detected by the use of ELISA, in particular where the immune effectors are detected using an ELISpot. 7. Postopek po zahtevku 1, kjer ima subjekt infekcijo s patogenim povzročiteljem, izbranim izmed Mycobacterium vrste, Staphylococcus vrste, Streptococcus vrste, Borrelia vrste, Escherichia coli, Salmonella vrste, Clostridium vrste, Shigella vrste, Proteus vrste, Bacillus vrste, Herpes virusa, virusa hepatitisa B ali C in virusa humane imunske pomanjkljivosti (HIV), ali bolezen, ki je rezultat le-tega, zlasti, kjer je bolezensko stanje infekcija z Mycobacterium tuberculosis ali tuberkuloza (TB), na primer, kjer je antigen izbran izmed CFP10, ESAT-6, TB7.7 in TB37.6.The method of claim 1 wherein the subject has an infection with a pathogenic agent selected from Mycobacterium species, Staphylococcus species, Streptococcus species, Borrelia species, Escherichia coli, Salmonella species, Clostridium species, Shigella species, Proteus species, Bacillus species, Herpes viruses, hepatitis B or C virus and human immunodeficiency virus (HIV) virus, or a disease resulting therefrom, in particular where the disease state is an infection with Mycobacterium tuberculosis or tuberculosis (TB), for example where the antigen is selected from CFP10, ESAT-6, TB7.7 and TB37.6. 8. Postopek po zahtevku 1, kjer ima subjekt bolezensko stanje, izbrano izmed naslednjih: alopecia areata, ankilozirajoči spondilitis, antifosfolipidni sindrom, autoimunska Addisonova bolezen, multipla skleroza, avtoimunska bolezen nadledvične žleze, avtoimunska hemolitična anemija, avtoimunski hepatitis, avtoimunski ooforitis in orhitis, Behcetova bolezen, bulozni pemfigoid, kardiomiopatija, celiakalična sprua-dermatitis, sindrom kronične utrujenosti (CFIDS), kronična vnetna demielinacija, kronična vnetna polinevropatija, Churg-Straussov sindrom, cikatrizacijski pemfigoid, crest sindrom, bolezen hladnega aglutinina, Crohnova bolezen, herpetiformen dermatitis, diskoidni lupus, esencialna mešana krioglobulinemija, fibromialgija, glomerulonefritis, Gravesova bolezen, Guillain-Barre, Hashimotov tiroiditis, idiopatska pljučna fibroza, idiopatska trombocitopenija purpura (ITP), IgA nefropatija, insulinsko odvisni diabetes (tip I), lichen planus, lupus, Menierova bolezen, mešana bolezen veznega tkiva, multipla skleroza, miastenija gravis, miokarditis, pemfigus vulgaris, pemiciozna anemija, nodozni poliarteriitis, polihondritis, poliglandulami sindromi, revmatična polimialgija, polimiozitis in dermatomiozitis, primarna agamaglobulinemija, primarna biliama ciroza, psoriaza, Raynaudov fenomen, Reiterjev sindrom, reumatična vročica, revmatoidni artritis, sarkoidoza, skleroderma, Sjogrenov sindrom, Stiff-Man sindrom, sistemski eritematozni lupus, Takayasujev arteriitis, temporalni arteriitis/ gigantocelulami arteriitis, ulcerozni kolitis, uveitis, vaskulitis, vitiligo in vnetna črevesna bolezen ali je bolezen celiakija ali je bolezen avtoimunski diabetes.The method of claim 1, wherein the subject has a disease state selected from: alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, multiple sclerosis, autoimmune adrenal gland disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and orchitis, Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac-sperm-dermatitis, chronic fatigue syndrome (CFIDS), chronic inflammatory demyelation, chronic inflammatory polyneuropathy, Churg-Strauss syndrome, cicatrization pemphigoid, crest syndrome, cold agglutinin disease, Crohn's disease, herpetiformen dermatitis, lupus, essential mixed cryoblobulinemia, fibromyalgia, glomerulonephritis, Graves disease, Guillain-Barre, Hashimoto thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP), IgA nephropathy, insulin dependent diabetes (type I), lichen planus, lupus, Menier's disease, it will be mixed connective tissue strap, multiple sclerosis, myasthenia gravis, myocarditis, pemphigus vulgaris, pemic anemia, nodosa polyarteritis, polycondritis, polyglandular syndromes, rheumatic polymyalgia, polymyositis and dermatomyositis, primary agamaglobulinemia, primary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever , rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, Stiff-Man syndrome, systemic lupus erythematosus, Takayasuva arteriitis, temporal arteritis / gigantocellular arteriitis, ulcerative colitis, uveitis, vasculitis, vitiligo and inflammatory bowel disease or is a disease of celiac disease or is autoimmune diabetes . 9. Postopek po zahtevku 1, kjer ima subjekt rak, izbran izmed naslednjih: ABL1 protoonkogen, z AIDS-om povezani raki, akustični nevrinom, akutna limfocitna levkemija, akutna mieloična levkemija, adenocistični karcinom, adrenokortikalni rak, agnogena mieloična metaplazija, alopecija, alveolami sarkom mehkih delov, analni rak, angiosarkom, aplastična anemija, astrocitom, ataksija-telangiektazija, bazalnocelični karcinom (koža), rak sečnega mehurja, kostni raki, črevesni rak, gliom možganskega debla, možganski in CŽS tumorji, rak dojk, SŽS tumoiji, karcinoidni tumorji, cervikalni rak, otroški možganski tumorji, otroški rak, otroška levkemija, otroški sarkom mehkega tkiva, hondrosarkom, horiokarcinom, kronična limfocitna levkemija, kronična mieloična levkemija, kolorektalni raki, kutani T-celični limfom, dermatofibrosarkom-protuberans, dezmoplastični-drobno-okroglo-celični-tumor, duktalni karcinom, endokrini raki, endometrijski rak, ependimom, ezofagealni rak, Ewingov sarkom, ekstrahepatični rak žolčevoda, očesni rak, očesni: melanom, retinoblastom, rak jajcevoda, Fanconijeva anemija, fibrosarkom, rak žolčnika, gastrični rak, gastrointestinalni raki, gastrointestinalni-karcinoidni-tumor, genitourinami raki, kličnocelični tumorji, gestacijska trofoblastna bolezen, gliom, ginekologoški raki, hematološke malignosti, lasnocelična levkemija, rak glave in vratu, hepatocelulami rak, hereditami rak dojke, histiocitoza, Hodgkinova bolezen, humani papilomavirus, hidatidozna mola, hiperkalcemija, rak hipofarinksa, intraokulami melanom, rak otočnih celic, Kaposijev sarkom, ledvični rak, histiocitoza Langerhansovih celic, laringealni rak, leiomiosarkom, levkemija, Li-Fraumenijev sindrom, ustnični rak, liposarkom, jetrni rak, pljučni rak, limfedem, limfom, Hodgkinov limfom, ne-Hodgkinov limfom, rak moških prsi, maligni rabdoidni tumor ledvice, meduloblastom, melanom, rak Merklovih celic, mezoteliom, metastatski rak, ustni rak, multipla endokrina neoplazija, fungoidna mikoza, mielodisplastični sindromi, mielom, mieloproliferativne motnje, nosni rak, nazofaringealni rak, nefroblastom, nevroblastom, nevrofibromatoza, sindrom lomov Nijmegen ("Nijmegen breackage syndrome"), ne-melanomski kožni rak, nedrobnocelični pljučni rak (NSCLC), očesni raki, ezofagealni rak, rak ustne votline, rak orofarinksa, osteosarkom, ostomijski ovarijski rak, pankreasni rak, paranazalni rak, paratiroidni rak, rak parotidne žleze, rak penisa, periferni nevroektodermalni tumoiji, hipofizni rak, prava policitemija, rak prostate, redki raki in povezane motnje, karcinom ledvičnih celic, retinoblastom, rabdomiosarkom, Rothmund-Thomsonov sindrom, rak žleze slinavke, sarkom, švanom, Sezaryjev sindrom, kožni rak, drobnocelični pljučni rak (SCLC), rak tankega črevesa, sarkom mehkega tkiva, tumorji hrbtenjače, ploščatocelični karcinom (koža), želodčni rak, sinovialni sarkom, testikulami rak, timusni rak, tiroidni rak, rak tranzicijskih celic (sečni mehur), rak tranzicijskih celic (ledvični-medenični-/-sečevoda), trofoblastični rak, rak sečevoda, rak urinamega sistema, uroplakini, sarkom maternice, rak maternice, vaginalni rak, rak vulve, Waldenstromova makroglobulinemija in Wilmsov tumor.The method of claim 1, wherein the cancer agent is selected from the following: ABL1 proto-oncogene, with AIDS-related cancer, acoustic neuron, acute lymphocytic leukemia, acute myeloid leukemia, adenocystic carcinoma, adrenocortical cancer, myeloid metaplasia agonist, alopecia, alveols soft tissue sarcoma, anal cancer, angiosarcoma, aplastic anemia, astrocytoma, ataxia-telangiectasia, basal cell carcinoma (skin), urinary bladder cancer, bone cancer, intestinal cancer, brain stem glioma, brain and CNS tumors, breast cancer, tumor tumor, carcinoid tumors, cervical cancer, child brain tumors, childhood cancer, childhood leukemia, soft tissue sock, chondrosarcoma, choriocarcinoma, chronic lymphocytic leukemia, chronic myeloid leukemia, colorectal cancer, corneal T-cell lymphoma, dermatophobrosarcoma-protuberance, desmoplastic-fine-round -cell-tumor, ductal carcinoma, endocrine cancer, endometrial cancer, ependymoma, esophageal cancer, Ewing gastric cancer, gastrointestinal cancer, gastrointestinal-carcinoma-tumor, genitourine cancer, klicocellular tumors, gestational trophoblastic disease, glioma, gastrointestinal disease, glioma, gastrointestinal disorders, glioma , gynecological cancer, haematological malignancies, lacosteal leukemia, head and neck cancer, hepatocellular cancer, hereditary breast cancer, histiocytosis, Hodgkin's disease, human papillomavirus, hydatidose mole, hypercalcemia, hypopharyngeal cancer, melanoma intraocular, ischemic cancer, Kaposi's sarcoma, kidney cancer , histiocytosis of Langerhans cells, laryngeal cancer, leiomyosarcoma, leukemia, Li-Fumumen syndrome, lipstick, liposarcoma, liver cancer, lung cancer, lymphedema, lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, male breast cancer, malignant rheumatoid tumor of the kidney, , melanoma, cancer of Merkel's cells, mesothelioma, metastatic cancer, oral cancer, multiple endocrine neuromuscular, neuroblastoma, neurosyptic, neuromuscular, neuropathy, neurosyphalitis, fracture syndrome, Nijmegen breackage syndrome, non-melanoma skin cancer, non-small cell lung cancer (NSCLC), nephrolithiasis, ocular cancer, esophageal cancer, oral cancer, oropharynx, osteosarcoma, ostomy ovarian cancer, pancreatic cancer, paranasal cancer, parathyroid cancer, parotid cancer, penile cancer, peripheral neuroectodermal tumors, pituitary cancer, real polycythemia, prostate cancer, rare cancers and related disorders, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, Rothmund-Thomson syndrome, salivary carcinoma, sarcoma, schwanum, Sezary syndrome, skin cancer, small cell lung cancer (SCLC), small intestine cancer, soft tissue sarcoma, spinal tumors, cancer (skin), gastric cancer, synovial sarcoma, cancer testicular cancer, thyroid cancer, thyroid cancer, transient cancer cerebrovascular cancer, uterine cancer, uterine cancer, uterine cancer, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, and Wilms tumor. 10. Postopek po zahtevku 1, kjer je bil subjekt izpostavljen proteinskemu toksikantu.The method of claim 1, wherein the subject has been exposed to a protein toxin. 11. Postopek po katerem koli od zahtevkov od 1 do 10, kjer magnituda celično posredovanega imunskega odziva korelira s stanjem, progresijo in/ali resnostjo bolezenskega stanja.The method of any one of claims 1 to 10, wherein the magnitude of the cell mediated immune response correlates with the condition, progression and / or severity of the disease state. 12. Uporaba dveh nizov peptidov, pri čemer prvi niz obsega peptide z dolžino od 7 do 14 aminokislinskih ostankov, ki jih prepoznavajo CD8+ limfociti, in drugi niz obsega peptide z od 16 aminokislinskih ostankov ali več, ki jih prepoznavajo CD4+ limfociti, pri čemer ti peptidi obsegajo ves ali del proteinskega antigena, v izdelavi diagnostičnega testa celično posredovane imunske odzivnosti s postopkom so-inkubacije kombinacije navedenih dveh nizov peptidov z limfociti in detektiranje prisotnosti ali zvišanja efektorskih molekul.Use of two strands of peptides, the first strand comprising peptides having a length of from 7 to 14 amino acid residues recognized by CD8 + lymphocytes, and the second strand comprising peptides having from 16 amino acid residues or higher recognized by CD4 + lymphocytes, wherein peptides comprise all or part of a protein antigen in the manufacture of a cell mediated immune response diagnostic test by co-incubation of a combination of said two strands of peptides with lymphocytes and detecting the presence or elevation of effector molecules. 13. Računalniško-izveden postopek, ki omogoča uporabniku, da določi status celično posredovane imuno-odzivnosti subjekta, pri čemer postopek obsega: (a) sprejemanje podatkov v obliki nivojev ali koncentracij imunske efektorske molekule, ki, glede na kontrolo, zagotovijo korelacijo s stanjem celično posredovane imuno-odzivnosti, od uporabnika preko komunikacijskega omrežja, pri čemer se imunsko efektorsko molekulo meri po so-inkubaciji limfocitov iz subjekta s kombinacijo dveh nizov peptidov, pri čemer prvi niz obsega peptide z dolžino od 7 do 14 aminokislinskih ostankov, ki jih prepoznavajo CD8+ limfociti, in drugi niz obsega peptide z od 16 aminokislinskih ostankov ali več, ki jih prepoznavajo CD4+ limfociti, pri čemer ti peptidi obsegajo ves ali del proteinskega antigena; (b) procesiranje subjektovih podatkov preko univariatne ali multivariatne analize, da se zagotovi vrednost imuno-odzivnosti; (c) določanje statusa subjekta skladno z rezultati vrednosti imuno-odzivnosti v primerjavi s predhodno določenimi vrednostmi; in (d) prenašanje indikacije statusa subjekta k uporabniku preko komunikacijskega omrežja.A computer-implemented procedure that allows the user to determine the status of the cell mediated immuno-responsiveness of the subject, the method comprising: (a) receiving data in the form of levels or concentrations of an immune effector molecule which, depending on the control, provide a correlation with the condition cell mediated immuno-responsiveness, from the user via the communication network, wherein the immune effector molecule is measured after co-incubation of the lymphocytes from the subject by a combination of two strands of peptides, the first strand comprising the peptides with a length of from 7 to 14 amino acid residues that they recognize CD8 + lymphocytes, and the second set comprises peptides having from 16 amino acid residues or more, recognized by CD4 + lymphocytes, these peptides comprising all or part of a protein antigen; (b) processing of subject data through a univariate or multivariate analysis to ensure the value of immuno-responsiveness; (c) determining the status of the subject in accordance with the results of the immuno-responsiveness value compared to the predetermined values; and (d) transmitting an indication of a subject's status to a user via a communication network.
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